| 摘要: |
| ［摘要］ 目的 研究人牙周膜干细胞（hPDLSCs）-条件培养液（CM）对MC3T3-E1细胞形态及增殖特性的影响。方法 从健康前磨牙及第三磨牙的牙周膜（牙周韧带）组织分离hPDLSCs并进行培养，采用免疫荧光法检测所得细胞中波形丝蛋白（vimentin）、角蛋白（PCK）及早期间充质干细胞标志物STRO-1的表达，鉴定细胞来源。制备hPDLSCs-CM对MC3T3-E1细胞进行干预处理，比较干预组与对照组（未经hPDLSCs-CM处理）MC3T3-E1细胞的形态特征和增殖能力。结果 免疫荧光染色显示，hPDLSCs呈现vimentin、STRO-1阳性表达、PCK阴性表达的特征。两组MC3T3-E1细胞均呈现长梭、多角的形态特征，贴壁生长。MTT实验结果显示，两组MC3T3-E1细胞的数量及增殖活性均随培养时间的增长呈现上升趋势。在培养第3天、第5天和第7天，干预组OD值均显著大于对照组，差异有统计学意义（P<0.05）。细胞周期分析结果显示，干预组和对照组处于G2/M+S期的细胞比例分别为(31.43±0.59)%和(19.11±0.24)%，比较差异有统计学意义（t=43.393，P=0.000）。结论 hPDLSCs-CM干预处理不影响MC3T3-E1细胞的形态特征，并可以增强其增殖能力。 |
| 关键词: 人牙周膜干细胞 条件培养液 MC3T3-E1细胞 细胞形态 增殖 |
| DOI：10.3969/j.issn.1674-3806.2021.02.09 |
| 分类号:R 787 |
| 基金项目:包头市医药卫生科技计划项目（编号：wsjj2019038） |
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| Effects of conditioned medium of human periodontal ligament stem cells on cell morphology and proliferative characteristics of mouse embryonic osteoblasts cells |
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LIU Yan, ZHAO Xiao-xia, WANG Jing-yu, et al.
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School of Stomatology, Baotou Medical College, Inner Mongolia 014060, China
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| Abstract: |
| ［Abstract］ Objective To study the effects of conditioned medium(CM) of human periodontal ligament stem cells(hPDLSCs) on cell morphology and proliferative characteristics of mouse embryonic osteoblasts(MC3T3-E1) cells. Methods The hPDLSCs were isolated from the pericementum(periodontal ligament) tissues of the healthy premolars and the third molars, and were cultured. Immunofluorescence was used to detect the expressions of vimentin, pan cytokeratin(PCK) and the early mesenchymal stem cell marker STRO-1 in the obtained cells to identify the source of the cells. The CM of hPDLSCs(hPDLSCs-CM) were prepared for the intervention treatment of MC3T3-E1 cells, and the morphological characteristics and proliferation ability of MC3T3-E1 cells were compared between the intervention group and the control group(without treatment of hPDLSCs-CM). Results Immunofluorescence staining showed that hPDLSCs showed the characteristics of vimentin and STRO-1 positive expressions, and PCK negative expression. The MC3T3-E1 cells in both groups showed the morphological characteristics of long spindles and polygons, and adherent growth. The results of methyl thiazolyl tetrazolium(MTT) experiment showed that the number and proliferation activity of MC3T3-E1 cells in the two groups showed an upward trend with the increase of culture time. On the 3rd, 5th and 7th day of culture, the OD value of the intervention group was significantly greater than that of the control group, and the difference was statistically significant(P<0.05). The results of cell cycle analysis showed that the proportion of cells in G2/M+S phase in the intervention group and the control group were (31.43±0.59)% and (19.11±0.24)%, respectively, and the difference was statistically significant(t=43.393,P=0.000). Conclusion Intervention with hPDLSCs-CM does not affect the morphological characteristics of MC3T3-E1 cells, and can enhance their proliferation ability. |
| Key words: Human periodontal ligament stem cells(hPDLSCs) Conditioned medium(CM) Mouse embryonic osteoblasts(MC3T3-E1) cells Cell morphology Proliferation |
